Biological pH buffers in IVF: help or hindrance to success

Will, M.; Clark, Natalie A.; Swain, Jason E.

doi:10.1007/s10815-011-9582-0

Cited by 95 publications

(51 citation statements)

References 102 publications

(191 reference statements)

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“…Analysis of pH revealed a significant difference in pH values of KSOMeq at 37°C in comparison with the the rest of treatments. These findings demonstrated that cryotubes allow CO 2 difussion into the air and a concomitant basification that was not buffered by the presence of HEPES as in KSOM-H, and that this diffusion was potentiated by temperature [17]. More efficient ways to preserve storage environment have been reported [18], that should be considered as an alternative if transport has to take place under such conditions.…”

Section: Resultsmentioning

confidence: 99%

In vitro and in vivodevelopment of mice morulae after storage in non-frozen conditions

Hourcade

Pérez-Crespo

Serrano

et al. 2012

Reprod Biol Endocrinol

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BackgroundInterchange of genetically modified (GM) mice between laboratories using embryos provides several advantages. Not only is transport stress avoided, but also the health status of the recipient colony is not compromised. Embryos do not need to be shipped in frozen stage, which requires expensive packaging in addition to a certain degree of expertise in order to freeze and thaw them correctly. The aim of this study was to examine different storage conditions and their effect on embryo viability in order to establish the feasibility of practical, non-frozen conditions for embryo shipment.MethodsMouse morulae developed in vivo (collected from donors 2.5d post coitum) or in vitro (zygotes cultured until morulae stage) were stored, combining two different media (KSOMeq or KSOM-H) and temperatures (4 degrees C, 15 degrees C and 37 degrees C) throughout 24 or 48 hours. After storage in vitro viability was assessed determining percentage of development to blastocyst and total cell number. In vivo viability was determined based on the number of implantations and living fetuses after embryo transfer of stored embryos. The storage effect at the molecular level was assessed by studying a gene pool involved in early development by quantitative RT-PCR.ResultsIn vivo-produced morulae stored for 24 hours did not show differences in development up to the blastocyst stage, regardless of the storage type. Even though a decrease in the total cell number in vivo was observed, embryo development after embryo transfer was not affected. All 24 hour storage conditions tested provided a similar number of implantations and fetuses at day 14 of pregnancy. Morulae obtained from in vitro embryo culture collected at the 1-cell stage showed a decreased ability to develop to blastocyst after 24 hours of storage at 15degrees C both in KSOMeq and KSOM-H. Concomitantly, a significant decrease of embryo implantation rates after transfer to recipients was also found. In order to further characterize the effect of non-frozen storage combining a molecular approach with the ordinary in vitro culture evaluation, embryos collected at the morula stage were submitted to the same storage conditions described throughout 48 hours. In vitro culture of those embryos showed a significant decrease in their developmental rate to blastocyst in both KSOMeq and KSOM-H at 15degrees C, which also affected the total number of cells. Gene transcription studies confirmed significant alterations in retrotransposons (Erv4 and Iap) after 48 h of storage at 15degrees C.ConclusionsOur results show that both KSOMeq and KSOM-H can be equally used, and that several temperature conditions allow good survival rates in vitro and in vivo. Some of these storage conditions can substitute freezing in order to maintain embryo viability for 24–48 hours, providing a reliable and less demanding technical alternative for embryo interchanges.

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Section: Resultsmentioning

confidence: 99%

In vitro and in vivodevelopment of mice morulae after storage in non-frozen conditions

Hourcade

Pérez-Crespo

Serrano

et al. 2012

Reprod Biol Endocrinol

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“…Thus, it is not unreasonable to conclude that ART intervention during the periconception period might induce a health risk in the resulting offspring. During ART, specifically in vitro gamete manipulation and embryo system, spermatozoa, oocytes, and embryos are exposed to various nonphysiological factors, such as culture media, altered pH [51], light [52,53], higher oxygen pressure [54], increased concentration of reproductive hormones [55], and mechanical injection or biopsy, all of which contribute to an altered periconception environment [56]. In addition to in vitro ART, a sub-optimal uterine environment following embryo transfer is thought to further influence the long-term viability of offspring.…”

Section: Discussionmentioning

confidence: 99%

Alteration in the expression of the renin-angiotensin system in the myocardium of mice conceived by in vitro fertilization†

Wang

Zhang

Fang

et al. 2018

Biology of Reproduction

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Epidemiological studies have revealed that offspring conceived by in vitro fertilization (IVF) have an elevated risk of cardiovascular malformations at birth, and are more predisposed to cardiovascular diseases. The renin-angiotensin system (RAS) plays an essential role in both the pathogenesis of congenital heart disease in fetuses and cardiovascular dysfunction in adults. This study aimed to assess the relative expression levels of genes in the RAS pathway in mice conceived using IVF, compared to natural mating with superovulation. Results demonstrated that expression of the angiotensin II receptor type 1 (AGTR1), connective tissue growth factor (CTGF), and collagen 3 (COL3), in the myocardial tissue of IVF-conceived mice, was elevated at 3 weeks, 10 weeks, and 1.5 years of age, when compared to their non-IVF counterparts. These data were supported by microRNA microarray analysis of the myocardial tissue of aged IVF-conceived mice, where miR-100, miR-297, and miR-758, which interact with COL3, AGTR1, and COL1 respectively, were upregulated when compared to naturally mated mice of the same age. Interestingly, bisulfite sequencing data indicated that IVF-conceived mice exhibited decreased methylation of CpG sites in Col1. In support of our in vivo investigations, miR-297 overexpression was shown to upregulate AGTR1 and CTGF, Effects of in vitro fertilization on myocardium, 2018, Vol. 99, No. 6 1277 and increased cell proliferation in cultured H9c2 cardiomyocytes. These findings indicate that the altered expression of RAS in myocardial tissue might contribute to cardiovascular malformation and/or dysfunction in IVF-conceived offspring. Furthermore, these cardiovascular abnormalities might be the result of altered DNA methylation and abnormal regulation of microRNAs. Summary SentenceAltered expression of the renin-angiotensin system in myocardial tissue might contribute to an increased risk of cardiovascular malformations and dysfunction in IVF offspring, and is involved in abnormal regulations of DNA methylation and microRNAs.

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“…There are many possible ways to improve our freezing system, such as changing the base medium from PBS to a 1:1 mixture of PBS and HEPES, as this mixture is known to stabilize pH better than PBS alone at low temperatures [37]. Antioxidants could also be added to the freezing and thawing media, as cryopreservation can generate reactive oxygen species that are harmful to cells [38].…”

Section: Discussionmentioning

confidence: 99%

Rapid freezing versus Cryotop vitrification of mouse two-cell embryos

Inna

Sanmee

Saeng-anan

et al. 2018

Clin Exp Reprod Med

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ObjectiveTo compare our in-house method of embryo freezing with Cryotop vitrification in terms of immediate survival, subsequent cleavage and blastocyst formation, and cell numbers in blastocysts.MethodsTwo-cell mouse embryos were randomly allocated into three groups: a non-frozen control group (group 1, n=300), a group that underwent Cryotop vitrification (group 2, n=300), and a group that underwent our in-house freezing method (group 3, n=300).ResultsThere were no significant differences between groups 2 and 3 in the immediate survival rate (96.3% vs. 98.6%, respectively; p=0.085), the further cleavage rate (91.7% vs. 95.0%, respectively; p=0.099), or the blastocyst formation rate (80.7% vs. 78.6%, respectively; p=0.437). The cell numbers in the blastocysts from groups 1, 2, and 3 were comparable (88.99±10.44, 88.29±14.79, and 86.42±15.23, respectively; p=0.228). However, the percentage of good-quality blastocysts in the Cryotop vitrification group was significantly higher than in the group in which our in-house method was performed, but was lower than in the control group (58.0%, 37.0%, and 82.7%, respectively; p<0.001).ConclusionAt present, our method is inferior to the commercial Cryotop vitrification system. However, with further improvements, it has the potential to be useful in routine practice, as it is easier to perform than the current vitrification system.

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Biological pH buffers in IVF: help or hindrance to success

Cited by 95 publications

References 102 publications

In vitro and in vivodevelopment of mice morulae after storage in non-frozen conditions

In vitro and in vivodevelopment of mice morulae after storage in non-frozen conditions

Alteration in the expression of the renin-angiotensin system in the myocardium of mice conceived by in vitro fertilization†

Rapid freezing versus Cryotop vitrification of mouse two-cell embryos

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